<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression

ABSTRACT The ectodomain of matrix protein 2 (M2e) of influenza A viruses is a universal influenza A vaccine candidate. Here, we report potential evasion strategies of influenza A viruses under in vivo passive anti-M2e IgG immune selection pressure in severe combined immune-deficient (SCID) mice. A/P...

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Autores principales:	Silvie Van den Hoecke, Marlies Ballegeer, Bram Vrancken, Lei Deng, Emma R. Job, Kenny Roose, Bert Schepens, Lien Van Hoecke, Philippe Lemey, Xavier Saelens
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	influenza broadly protective deep sequencing escape immunotherapy monoclonal antibodies Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d51386aa0cb244889e4bdf58c40d32b6
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spelling	oai:doaj.org-article:d51386aa0cb244889e4bdf58c40d32b62021-11-10T18:37:50Z<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression10.1128/mBio.00745-212150-7511https://doaj.org/article/d51386aa0cb244889e4bdf58c40d32b62021-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00745-21https://doaj.org/toc/2150-7511ABSTRACT The ectodomain of matrix protein 2 (M2e) of influenza A viruses is a universal influenza A vaccine candidate. Here, we report potential evasion strategies of influenza A viruses under in vivo passive anti-M2e IgG immune selection pressure in severe combined immune-deficient (SCID) mice. A/Puerto Rico/8/34-infected SCID mice were treated with the M2e-specific mouse IgG monoclonal antibodies (MAbs) MAb 65 (IgG2a) or MAb 37 (IgG1), which recognize amino acids 5 to 15 in M2e, or with MAb 148 (IgG1), which binds to the invariant N terminus of M2e. Treatment of challenged SCID mice with any of these MAbs significantly prolonged survival compared to isotype control IgG treatment. Furthermore, M2e-specific IgG2a protected significantly better than IgG1, and even resulted in virus clearance in some of the SCID mice. Deep sequencing analysis of viral RNA isolated at different time points after treatment revealed that the sequence variation in M2e was limited to P10H/L and/or I11T in anti-M2e MAb-treated mice. Remarkably, in half of the samples isolated from moribund MAb 37-treated mice and in all MAb 148-treated mice, virus was isolated with a wild-type M2 sequence but with nonsynonymous mutations in the polymerases and/or the hemagglutinin genes. Some of these mutations were associated with delayed M2 and other viral gene expression and with increased resistance to anti-M2e MAb treatment of SCID mice. Treatment with M2e-specific MAbs thus selects for viruses with limited variation in M2e. Importantly, influenza A viruses may also undergo an alternative escape route by acquiring mutations that result in delayed wild-type M2 expression. IMPORTANCE Broadly protective influenza vaccine candidates may have a higher barrier to immune evasion compared to conventional influenza vaccines. We used Illumina MiSeq deep sequence analysis to study the mutational patterns in A/Puerto Rico/8/34 viruses that evolve in chronically infected SCID mice that were treated with different M2e-specific MAbs. We show that under these circumstances, viruses emerged in vivo with mutations in M2e that were limited to positions 10 and 11. Moreover, we discovered an alternative route for anti-M2e antibody immune escape, in which a virus is selected with wild-type M2e but with mutations in other gene segments that result in delayed M2 and other viral protein expression. Delayed expression of the viral antigen that is targeted by a protective antibody thus represents an influenza virus immune escape mechanism that does not involve epitope alterations.Silvie Van den HoeckeMarlies BallegeerBram VranckenLei DengEmma R. JobKenny RooseBert SchepensLien Van HoeckePhilippe LemeyXavier SaelensAmerican Society for Microbiologyarticleinfluenzabroadly protectivedeep sequencingescapeimmunotherapymonoclonal antibodiesMicrobiologyQR1-502ENmBio, Vol 12, Iss 4 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	influenza broadly protective deep sequencing escape immunotherapy monoclonal antibodies Microbiology QR1-502
spellingShingle	influenza broadly protective deep sequencing escape immunotherapy monoclonal antibodies Microbiology QR1-502 Silvie Van den Hoecke Marlies Ballegeer Bram Vrancken Lei Deng Emma R. Job Kenny Roose Bert Schepens Lien Van Hoecke Philippe Lemey Xavier Saelens <italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
description	ABSTRACT The ectodomain of matrix protein 2 (M2e) of influenza A viruses is a universal influenza A vaccine candidate. Here, we report potential evasion strategies of influenza A viruses under in vivo passive anti-M2e IgG immune selection pressure in severe combined immune-deficient (SCID) mice. A/Puerto Rico/8/34-infected SCID mice were treated with the M2e-specific mouse IgG monoclonal antibodies (MAbs) MAb 65 (IgG2a) or MAb 37 (IgG1), which recognize amino acids 5 to 15 in M2e, or with MAb 148 (IgG1), which binds to the invariant N terminus of M2e. Treatment of challenged SCID mice with any of these MAbs significantly prolonged survival compared to isotype control IgG treatment. Furthermore, M2e-specific IgG2a protected significantly better than IgG1, and even resulted in virus clearance in some of the SCID mice. Deep sequencing analysis of viral RNA isolated at different time points after treatment revealed that the sequence variation in M2e was limited to P10H/L and/or I11T in anti-M2e MAb-treated mice. Remarkably, in half of the samples isolated from moribund MAb 37-treated mice and in all MAb 148-treated mice, virus was isolated with a wild-type M2 sequence but with nonsynonymous mutations in the polymerases and/or the hemagglutinin genes. Some of these mutations were associated with delayed M2 and other viral gene expression and with increased resistance to anti-M2e MAb treatment of SCID mice. Treatment with M2e-specific MAbs thus selects for viruses with limited variation in M2e. Importantly, influenza A viruses may also undergo an alternative escape route by acquiring mutations that result in delayed wild-type M2 expression. IMPORTANCE Broadly protective influenza vaccine candidates may have a higher barrier to immune evasion compared to conventional influenza vaccines. We used Illumina MiSeq deep sequence analysis to study the mutational patterns in A/Puerto Rico/8/34 viruses that evolve in chronically infected SCID mice that were treated with different M2e-specific MAbs. We show that under these circumstances, viruses emerged in vivo with mutations in M2e that were limited to positions 10 and 11. Moreover, we discovered an alternative route for anti-M2e antibody immune escape, in which a virus is selected with wild-type M2e but with mutations in other gene segments that result in delayed M2 and other viral protein expression. Delayed expression of the viral antigen that is targeted by a protective antibody thus represents an influenza virus immune escape mechanism that does not involve epitope alterations.
format	article
author	Silvie Van den Hoecke Marlies Ballegeer Bram Vrancken Lei Deng Emma R. Job Kenny Roose Bert Schepens Lien Van Hoecke Philippe Lemey Xavier Saelens
author_facet	Silvie Van den Hoecke Marlies Ballegeer Bram Vrancken Lei Deng Emma R. Job Kenny Roose Bert Schepens Lien Van Hoecke Philippe Lemey Xavier Saelens
author_sort	Silvie Van den Hoecke
title	<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
title_short	<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
title_full	<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
title_fullStr	<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
title_full_unstemmed	<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression
title_sort	<italic toggle="yes">in vivo</italic> therapy with m2e-specific igg selects for an influenza a virus mutant with delayed matrix protein 2 expression
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/d51386aa0cb244889e4bdf58c40d32b6
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<italic toggle="yes">In Vivo</italic> Therapy with M2e-Specific IgG Selects for an Influenza A Virus Mutant with Delayed Matrix Protein 2 Expression

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